Large scale lentiviral vector production

Abstract

Lentiviral vectors (LV) are remarkable in their ability to insert their genetic payload into a target cell's genome. This affects apermanent genetic change in the target which is propagated through to its progeny. While there are several means ofgenetic modification, few are capable of permanent modification of target cells. Amongst all vectors for gene delivery, LVare unique in unparalleled efficiency, safety, lack of toxicity and ability to modify non-dividing target cells. They havetherefore come to be recognised as a key reagent required for the efficient development of the burgeoning cell therapy, aswell as gene therapy industries. However, although LV represent a well understood and robust technology, there is nomanufacturing methodology for very large-scale LV production. This is now an acknowledged bottle-neck both for clinicaltrials and for commercial exploitation of many cell and gene therapy products in current development. We propose toaddress this unmet need

Technical Summary

* Lentiviral vectors (LV) are used to genetically modify target cells. The main commercial application of LVs is in vitroproduction of cellular therapeutics, although they can be used as therapeutic agents in their own right and for generation ofprotein producer cells. While small-scale production is well established, a process for very large-scale production has notbeen developed. Recently, commercial demand for LV has increased greatly. Large-scale production is a major unmetneed and its lack a bottleneck esp. in the marketing of cellular therapies. The challenge of large-scale LV production ismultifaceted, requiring a deep understanding of lentiviral biology, as well as skilful genetic and cellular engineering,appreciation of regulatory / GMP considerations, industrial/pharmaceutical scale production issues as well as anunderstanding of the down-stream clinical applications.

The single most important contribution of this project has been effective process development and manufacture of reagents (cells and vectors) needed for the manufacture of gamma-retrovirus and lentivirus vectors that are used for various clinical applications of cell and gene therapy.

Exploitation Route

The findings of two previous BBSRC grants (BB/E005896/1 and BB/D014301/1) and the current project (BB/N003853/1) and to a smaller extent BB/K013785/1, have underpinned the development of a major manufacturing activity (GMP Production of viral vectors for clinical use) at King's College London. We are now in active collaborations with a number of pharmaceutical (Cellectis, as well as Pfizer and Servier in active discussions for further collaborations) and biotech companies (Autolus and LiFT), as well as the Cell Therapy Catapult, for the further development and commercial exploitation of these manufacturing facilities and experties, including IP.

This research, in combination with BB/K013785/1 and BB/E005896/1 has underpinned a great deal of innovation in the manufacture and production of Lentivirus and retrovirus vectors for clinical trials. This IP, combined with a GLP/GMP facility supported by grants from NIHR and CRUK have enabled the attraction of a number of major research grants including £2.5M from Roche Pharmaceuticals for the evaluation of glycoengineered antibodies and the evaluation of immunological markers in patients receiving these antibodies in phase-i/II clinicaal trials. The IP generated, the facilities and the experience of working on the Roche supported projects have in turn resulted in the attraction of manufacturing contracts from a US Biotech company (Northwest Bio - 1.2M), from a French company focused on the generation of allogeneic T cells expressing Chimeric antigen Receptors (Cellectis, over £11M between 2016 and 2019). The contract with this compnay, Cellectis, is worth in total about £15M over the next 3 years (2016-2019). We are also engaged in a fruther collaborations with a new biotech company in UK (Autolous) for the production of retroviral vectors that are used in the generation of other CAR-T cells for clinical studies. Finally, a recently initiated collaboration with Cell and Gene Therapy Catapult is developing procedures and products for the industrialisation of cell and gene therapy products, including the development of a scalable strategy for suspension cell based manufacture of retrovirus vectors encoding specific T cell receptors.

Cellectis: Production of viral vectors (primarily lentivirus), and gene modified cells, for clinical applications of cell and gene therapy

Organisation

Cellectis

Country

France

Sector

Private

PI Contribution

Development and production of multiple lentivirus and retrovirus vectors for a range of clinical studies in collaboration with both academic and industry partners, the largest of which is the collaboration with Cellectis culminating in over £1.6 million of funding todate, plus a new contract for £10.1 million over the next 3 years.

Collaborator Contribution

Provision of funding and know-how in specific areas (e.g. site directed endonuclease mediated inhibition of endogenous T cell receptors, in order to allow the generation of allogeneic (off-the-shelf) Chimeric Antigen Receptor (CAR) T cells for the treatment of malignant disease. This project is directly supported by BB/N003853/1 and assisted by the outputs from our previous BBSRC grants: BB/E005896/1, BB/D014301/1 and BB/K013785/1.

Impact

The development of allogeneic CAR-T cells (referred to as UniCAR-T) for the treatment of malignant disease. There has also been substantial inward investment (over £11,000,000 between 2016 and 2019 from Cellectis alone) underpining further developments that we expect to culminate in substantially larger collaborations with other pharmaceuticaal companies (active discussions in progress with Cell Therapy Catapult, Pfizer and Servier). This collaboration has also resulted in a separate collaboration with a UK based start-up company - Autolus (reported as a separate collaboration).

Start Year

2015

Description

Collaboration with the University fo Lausanne for the development and production of lentivirus lectors

Organisation

University of Lausanne

Country

Switzerland

Sector

Academic/University

PI Contribution

We have set up a new collaboration with the University of Lausanne for the development of lentivirus vectors and their GMP manufacture over the next 3 years (2018 to 2020). University of Lausanne has provided a contract of £2.6M of which the first instalment of 20% has already been paid.

Collaborator Contribution

The development of vectors that they have produced for a number of gene therapy based clinical studies, and the use of the vectors made under GMP at King's College London, in these clinical trials.

Impact

Contracts of Collaboration Signed

Start Year

2018

Description

Vector Industrialisation Project

Organisation

Cell Therapy Catapult

Country

United Kingdom

Sector

Academic/University

PI Contribution

The aim of this recently initiated project is the development of GMP compatible procedures for the industrialisation of gene therapy products. The main focus of this specific project is the development of cell line/s with characteristics needed for large scale manufacture of a retrovirus vector encoding a specific T cell receptor (TCR).

Collaborator Contribution

Funding of the initial studies, providing the vector manufacturing cell line and expertise in project management, accurate calculation of costs of goods. Cell and Gene Therapy Catapult is also providing expertise in the development of strategies aimed at reducing the cost of goods, risk-reduction for manufacturing campaigns and strategies for efficient large scale manufacture of clinical grade vectors. This project is underpinned by two previous BBSRC grants and directly affected by our current BBSRC supported project.

Impact

This project is contributing to the development of new therapies and therapeutic strategies, particularly with respect of the industrial scale manufacture of cell and gene therapy vectors, thus contributing both to better health care and to creation of wealth, including inward investment from outside the UK.

Start Year

2016

Title

Development of GMP compliant manufacturing strategies for the production of clinical grade viral vectors

Description

The production of viral vectors, in particular lentivirus and gamma-retrovirus in sufficient quantities and able to meet the regulatory standards of quality is particularly challenging. Using the technologies that were developed as part of our BBSRC supported projects, we have established a range of manufacturing, purification and concentration strategies that have enabled us to manufacture the largest number (academia or industry) of retroviral and lentivirus vectors for regulatory approved clinical trials in Europe. This extensive research and development programme has now culminated in over £15 million pound of income (2012 to 2019) for King's College London from overseas based companies.

IP Reference

Protection

Protection not required

Year Protection Granted

2016

Licensed

Yes

Impact

The background manufacturing IP and know-how is licensed (non-exclusive) to Cellectis and to Cell Therapy Catapult. Discussions are in progress with other organisations in taking similar non-exclusive licenses.

Title

The processes developed in the course of this study have directly contributed to the success of subsequent contracts with the Industry, including Autolus and Cellectis (biotech and pharmaceutical companies.

Description

We have developed procedures for the fast manufacture of retrovirus and lentivirus vectors in compliance with the regulatory requirements for clinical use (GMP compliant procedures). These highly optimised procedures have enabled the production of high titre vectors (about 50,000 million infectious units of vector) from relatively small scale cultures (circa 10 litres), with greater than 50% recovery (frequently in excess of 70%) and minimal quantities of contaminating proteins and nucleic acids. This knowhow has recently been licensed on non-exclusive deals to the industry (Cellectis) in contracts producing in excess of £15 million pounds of income over the next 3 years.

IP Reference

Protection

Protection not required

Year Protection Granted

2016

Licensed

Yes

Impact

We have produced, for regulatory approved clinical trials, the largest number of lenti- and retroviral vectors in Europe. Each of the 4 BBSRC supported projects have contributed to this outcome. We are now extending this expertise with a view to similarly innovative manufacture of Adeno Associated Virus (AAV) manufacture for clinical use.

Title

CAR T cells

Description

The GMP vectors produced with the support of this BBSRC grant are now being used in a range of clinical trials based at Uuniversity College Hospital, Great Ormod Street Hospital and King's College Hospital

Type

Therapeutic Intervention - Cellular and gene therapies

Current Stage Of Development

Early clinical assessment

Year Development Stage Completed

2016

Development Status

Under active development/distribution

Clinical Trial?

Yes

Impact

The vectors produced with the support of this project are now being used in multiple clinical trials - all Phase-1, all first-in-man

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